Evaluation of the antiviral-HAdV activity of venom extracts obtained from arachnids and snakes.

Motivation: Human adenoviruses (HAdVs) can cause life-threatening infections and disseminated disease in hematopoietic stem cell transplant (HSCT) recipients, especially in paediatric patients, and is also a well-known cause of community-acquired pneumonia in the general population. Despite this significant impact on public health, there are no specific antiviral therapies approved by the health authorities to treat HAdV infections. Thus, it is necessary to identify and characterize molecules that exert their antiviral action against HAdVs infection as the first step in the clinical development of an specific antiviral drug. In this sense, we have selected a collection of crude extracts of arachnid and snake venoms to evaluate their antiviral activity against HAdV.Methods: A total of 67 extracts from different arachnid species were subjected to a first screening at a concentration of 50 μg/ml in A549 cells infected with HAdVs-RFP at a MOI of 2000 vp/cell for 48 h. Those that showed an inhibition of the viral infection higher than 60% were tested through the same procedure using HAdV-GFP to determine if the extract exerts its antiviral action in early or late stages of the HAdV replicative cycle. In parallel, the selected compounds were subjected to a methabolic assay to determine their cytotoxicity (CC50). Finally, the selected compounds were tested at a low MOI (0.06 vp/cell) with HAdV-GFP at concentrations ranging from 10 μg/ml to 0,625 μg/ml. For the extracts that their inhibitory concentration 50% (IC50) could be calculated, the selectivity index (SI) was determined, as the coefficient between the CC50 and the IC50.Results: Sixteen out of the 67 extracts analyzed with HAdVs-RFP and HAdV-GFP, showed an inhibition of the infection >60%. The CC50 values obtained ranged from 87.43 ± 27.86 to 1.24 ± 0.21 μg/ml. So far, most of the extracts evaluated present a IC50 value above 10 μg/ml, with the exception of Bothrops atrox and Bothrops pictus, with an IC50 value of 0.75 ± 0.047 μg/ml (SI = 24,26) and 1.3 ± 0.00565 μg/ml (SI = 23,20), respectively.Conclusions: Sixteen extracts have been identified with the ability to inhibit HAdV infection by more than 60%, and presenting CC50 above their IC50. Thus, in this situation, the next step will be to carry out the deconvolution of these 16 extracts, to identify the molecule/s responsible for the observed activity in order to accurately characterize their antiviral activity and mechanism of action

Autoantibodies against type I IFNs in patients with critical influenza pneumonia

In an international cohort of 279 patients with hypoxemic influenza pneumonia, we identified 13 patients (4.6%) with autoantibodies neutralizing IFN-alpha and/or -omega, which were previously reported to underlie 15% cases of life-threatening COVID-19 pneumonia and one third of severe adverse reactions to live-attenuated yellow fever vaccine. Autoantibodies neutralizing type I interferons (IFNs) can underlie critical COVID-19 pneumonia and yellow fever vaccine disease. We report here on 13 patients harboring autoantibodies neutralizing IFN-alpha 2 alone (five patients) or with IFN-omega (eight patients) from a cohort of 279 patients (4.7%) aged 6-73 yr with critical influenza pneumonia. Nine and four patients had antibodies neutralizing high and low concentrations, respectively, of IFN-alpha 2, and six and two patients had antibodies neutralizing high and low concentrations, respectively, of IFN-omega. The patients' autoantibodies increased influenza A virus replication in both A549 cells and reconstituted human airway epithelia. The prevalence of these antibodies was significantly higher than that in the general population for patients 70 yr of age (3.1 vs. 4.4%, P = 0.68). The risk of critical influenza was highest in patients with antibodies neutralizing high concentrations of both IFN-alpha 2 and IFN-omega (OR = 11.7, P = 1.3 x 10(-5)), especially those <70 yr old (OR = 139.9, P = 3.1 x 10(-10)). We also identified 10 patients in additional influenza patient cohorts. Autoantibodies neutralizing type I IFNs account for similar to 5% of cases of life-threatening influenza pneumonia in patients <70 yr old

Dysregulation in host sphingolipid metabolism caused by West Nile virus infection as a potential disease progression biomarker and new therapeutic target

West Nile virus (WNV) is a neurotropic flavivirus transmitted by the bites of infected Culex sp mosquitoes and it is widely distributed across Europe. Severe forms of West Nile disease (WND) can cause meningitis, encephalitis or even death. In 2020, WNV caused an outbreak in southwest Spain that ended up with 70 human infections and 7 deaths. There are no vaccines or specific drugs to prevent and treat the symptoms of WND in humans, so the search for novel targets for therapeutic interventions is mandatory. Lipid metabolism is gaining interest as a potential target to combat flavivirus using a host-targeted approach.Peer reviewe

Concomitant administration of seasonal influenza and COVID-19 mRNA vaccines

ABSTRACTCurrent clinical guidelines support the concomitant administration of seasonal influenza vaccines and COVID-19 mRNA boosters vaccine. Whether dual vaccination may impact vaccine immunogenicity due to an interference between influenza or SARS-CoV-2 antigens is unknown. We aimed to understand the impact of mRNA COVID-19 vaccines administered concomitantly on the immune response to influenza vaccines. For this, 128 volunteers were vaccinated during the 22-23 influenza season. Three groups of vaccination were assembled: FLU vaccine only (46, 35%) versus volunteers that received the mRNA bivalent COVID-19 vaccines concomitantly to seasonal influenza vaccines, FluCOVID vaccine in the same arm (42, 33%) or different arm (40, 31%), respectively. Sera and whole blood were obtained the day of vaccination, +7, and +28 days after for antibody and T cells response quantification. As expected, side effects were increased in individuals who received the FluCOVID vaccine as compared to FLU vaccine only based on the known reactogenicity of mRNA vaccines. In general, antibody levels were high at 4 weeks post-vaccination and differences were found only for the H3N2 virus when administered in different arms compared to the other groups at day 28 post-vaccination. Additionally, our data showed that subjects that received the FluCOVID vaccine in different arm tended to have better antibody induction than those receiving FLU vaccines for H3N2 virus in the absence of pre-existing immunity. Furthermore, no notable differences in the influenza-specific cellular immune response were found for any of the vaccination groups. Our data supports the concomitant administration of seasonal influenza and mRNA COVID-19 vaccines

DataSheet1_Inhibition of adenovirus transport from the endosome to the cell nucleus by rotenone.pdf

Regardless of the clinical impact of human adenovirus (HAdV) infections in the healthy population and its high morbidity in immunosuppressed patients, a specific treatment is still not yet available. In this study, we screened the CM1407 COST Action’s chemical library, comprising 1,233 natural products to identify compounds that restrict HAdV infection. Among them, we identified rotenolone, a compound that significantly inhibited HAdV infection. Next, we selected four isoflavonoid-type compounds (e.g., rotenone, deguelin, millettone, and tephrosin), namely rotenoids, structurally related to rotenolone in order to evaluate and characterized in vitro their antiviral activities against HAdV and human cytomegalovirus (HCMV). Their IC50 values for HAdV ranged from 0.0039 µM for rotenone to 0.07 µM for tephrosin, with selective indices ranging from 164.1 for rotenone to 2,429.3 for deguelin. In addition, the inhibition of HCMV replication ranged from 50% to 92.1% at twice the IC50 concentrations obtained in the plaque assay for each compound against HAdV. Our results indicated that the mechanisms of action of rotenolone, deguelin, and tephrosin involve the late stages of the HAdV replication cycle. However, the antiviral mechanism of action of rotenone appears to involve the alteration of the microtubular polymerization, which prevents HAdV particles from reaching the nuclear membrane of the cell. These isoflavonoid-type compounds exert high antiviral activity against HAdV at nanomolar concentrations, and can be considered strong hit candidates for the development of a new class of broad-spectrum antiviral drugs.</p

Lipid signatures of West Nile virus infection unveil alterations of sphingolipid metabolism providing novel biomarkers

AbstractWest Nile virus (WNV) is a neurotropic flavivirus transmitted by the bites of infected mosquitoes. Severe forms of West Nile disease (WND) can curse with meningitis, encephalitis or acute flaccid paralysis. A better understanding of the physiopathology associated with disease progression is mandatory to find biomarkers and effective therapies. In this scenario, blood derivatives (plasma and serum) constitute the more commonly used biofluids due to its ease of collection and high value for diagnostic purposes. Therefore, the potential impact of this virus in the circulating lipidome was addressed combining the analysis of samples from experimentally infected mice and naturally WND patients. Our results unveil dynamic alterations in the lipidome that define specific metabolic fingerprints of different infection stages. Concomitant with neuroinvasion in mice, the lipid landscape was dominated by a metabolic reprograming that resulted in significant elevations of circulating sphingolipids (ceramides, dihydroceramides and dihydrosphingomyelins), phosphatidylethanolamines and triacylglycerols. Remarkably, patients suffering from WND also displayed an elevation of ceramides, dihydroceramides, lactosylceramides and monoacylglycerols in their sera. The dysregulation of sphingolipid metabolism by WNV may provide new therapeutic opportunities and supports the potential of certain lipids as novel peripheral biomarkers of WND progression

Concomitant administration of seasonal influenza and COVID-19 mRNA vaccines

Current clinical guidelines support the concomitant administration of seasonal influenza vaccines and COVID-19 mRNA boosters vaccine. Whether dual vaccination may impact vaccine immunogenicity due to an interference between influenza or SARS-CoV-2 antigens is unknown. We aimed to understand the impact of mRNA COVID-19 vaccines administered concomitantly on the immune response to influenza vaccines. For this, 128 volunteers were vaccinated during the 22-23 influenza season. Three groups of vaccination were assembled: FLU vaccine only (46, 35%) versus volunteers that received the mRNA bivalent COVID-19 vaccines concomitantly to seasonal influenza vaccines, FluCOVID vaccine in the same arm (42, 33%) or different arm (40, 31%), respectively. Sera and whole blood were obtained the day of vaccination, +7, and +28 days after for antibody and T cells response quantification. As expected, side effects were increased in individuals who received the FluCOVID vaccine as compared to FLU vaccine only based on the known reactogenicity of mRNA vaccines. In general, antibody levels were high at 4 weeks post-vaccination and differences were found only for the H3N2 virus when administered in different arms compared to the other groups at day 28 post-vaccination. Additionally, our data showed that subjects that received the FluCOVID vaccine in different arm tended to have better antibody induction than those receiving FLU vaccines for H3N2 virus in the absence of pre-existing immunity. Furthermore, no notable differences in the influenza-specific cellular immune response were found for any of the vaccination groups. Our data supports the concomitant administration of seasonal influenza and mRNA COVID-19 vaccines.</p

Autoantibodies against type I IFNs in patients with critical influenza pneumonia

International audienceAutoantibodies neutralizing type I interferons (IFNs) can underlie critical COVID-19 pneumonia and yellow fever vaccine disease. We report here on 13 patients harboring autoantibodies neutralizing IFN-α2 alone (five patients) or with IFN-ω (eight patients) from a cohort of 279 patients (4.7%) aged 6–73 yr with critical influenza pneumonia. Nine and four patients had antibodies neutralizing high and low concentrations, respectively, of IFN-α2, and six and two patients had antibodies neutralizing high and low concentrations, respectively, of IFN-ω. The patients’ autoantibodies increased influenza A virus replication in both A549 cells and reconstituted human airway epithelia. The prevalence of these antibodies was significantly higher than that in the general population for patients &lt;70 yr of age (5.7 vs. 1.1%, P = 2.2 × 10−5), but not &gt;70 yr of age (3.1 vs. 4.4%, P = 0.68). The risk of critical influenza was highest in patients with antibodies neutralizing high concentrations of both IFN-α2 and IFN-ω (OR = 11.7, P = 1.3 × 10−5), especially those &lt;70 yr old (OR = 139.9, P = 3.1 × 10−10). We also identified 10 patients in additional influenza patient cohorts. Autoantibodies neutralizing type I IFNs account for ∼5% of cases of life-threatening influenza pneumonia in patients &lt;70 yr old